In this project we propose to build a multi-disciplinary and multi-technological platform to address the phenotypic, functional, and molecular characterization of genetic modified mice. This is of utility for translating the information stored in the human genome into increasingly accurate models of human disease. The "Platform", consisting of different experimental units, such as morphology, functional & behavioral, molecular biology, metabolic, microsurgery, and diagnostic fertility unit, is designed to assist the researcher in dealing with the characterization of the mouse models, through specific analysis and standardized procedures. Our project aims to standardize the phenotyping procedures and to assist investigators in their experimental design and execution by providing an extensive baseline phenotypic profile of mice and specific morphological, functional and molecular analysis. This novel platform enables also preclinical research on primary defects in order to test new treatment options. In this context, the platform provides investigators with a central high-quality microsurgical service to design and develop models that require microsurgery in small animals that allow investigators to test their hypothesis.
Of note, most of the "hubs" related to the platform (functional unit, performance and metabolic unit, molecular biology unit, microsurgery unit, diagnostic fertility unit) are already activated and equipped. We would like to expand and implement the morphology unit for the analysis of immunophenotyping of complex structures. Several approaches can be proposed, according to our expertise. This platform intends to capitalize the investment already made in the generation of several mouse models, offering to the research community the opportunity to analyse the models at different and professional levels.
Genotype-phenotype studies aim to identify causative relationships between genes and phenotypes.
High-throughput phenotyping is a cornerstone of numerous functional genomics projects. In recent years, imaging screens have become increasingly important in understanding gene-phenotype relationships in studies of cells, tissues and whole organisms. Three-dimensional (3D) imaging has risen to prominence in the field of developmental biology for its ability to capture whole embryo morphology and gene expression.
Understanding the relationship of the structure of organs to their function is a key component of integrative physiological research. The structure of the organs of the body is not constant but changes, both during growth and development and under conditions of sustained stress. Recently, powerful new techniques have become available in molecular biology, which promise to provide novel insights into the mechanisms and consequences of these altered structure-function relationships.
In this project we plan to use, along conventional technical approaches, strategy to analyse complex structure, such as neuromuscular junction and to define their three-dimensional organization.
Conventionally structure-function relationships are studied by microscopic examination of tissue sections. However, drawing conclusions about the three-dimensional structure of an organ based on this two-dimensional information frequently leads to serious errors. The techniques of stereology allow precise and accurate quantification of structural features within three-dimensional organs that relate in a meaningful way to integrated function. For example, knowledge of changes in the total surface area of the capillary endothelium in a n organ can be related directly to changes in fluid filtration and permeability, or knowledge of total vessel length and mean radius allows deductions about vascular resistance. Confocal microscopy adds enormously to the power of stereological approaches. It reduces the difficulties and labour involved in obtaining suitable images. Moreover, when used in conjunction with new analytical software, it allows convenient application of stereology to small samples and those in which it is essential to maintain a specific orientation for interpretation. The information obtained will allow us to examine in a quantitative manner the altered structure-function relationships produced by manipulation of single genes and regulatory pathways in whole organisms.
Laser scanning confocal microscopy is a significant advance in the field of optical microscopy, primarily because it permits sample visualization deep within living and fixed cells, tissues and other samples. It provides the ability to collect sharply defined optical sections from which three-dimensional renderings can be created.
Thus, we propose to build a multi-disciplinary and multi-technological platform to perform phenotypic, functional, molecular and behavioral test on experimental models.
The "Platform", consisting of different experimental units is designed to assist the researcher in dealing with the characterization of the mouse models, through specific analysis and standardized procedures. This novel platform enables also preclinical research on primary defects in order to test new treatment options. In this context, the platform provides investigators with a central high-quality service to design and develop models that require microsurgery in small animals that allow investigators to test their hypothesis.